characterization of cardiac stress (takotsubo cardiomyopathy)-related mirnas ex vivo anselm derda,...
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![Page 1: Characterization of cardiac stress (Takotsubo Cardiomyopathy)-related miRNAs ex vivo Anselm Derda, ESC Congress 2015 NHLI – Prof. Sian Harding IMTTS –](https://reader035.vdocuments.site/reader035/viewer/2022062321/56649f355503460f94c527e9/html5/thumbnails/1.jpg)
Characterization of cardiac stress (Takotsubo Cardiomyopathy)-related miRNAs ex vivo
Anselm Derda, ESC Congress 2015NHLI – Prof. Sian Harding
IMTTS – Prof. Thomas Thum
![Page 2: Characterization of cardiac stress (Takotsubo Cardiomyopathy)-related miRNAs ex vivo Anselm Derda, ESC Congress 2015 NHLI – Prof. Sian Harding IMTTS –](https://reader035.vdocuments.site/reader035/viewer/2022062321/56649f355503460f94c527e9/html5/thumbnails/2.jpg)
1. 2. 3.
Isolation of LV rat myocytes
Transfection & Incubation for 48h Control/Inhibitor/Mimic of miR-16
Contractility/IonOptix
Basal contractility
![Page 3: Characterization of cardiac stress (Takotsubo Cardiomyopathy)-related miRNAs ex vivo Anselm Derda, ESC Congress 2015 NHLI – Prof. Sian Harding IMTTS –](https://reader035.vdocuments.site/reader035/viewer/2022062321/56649f355503460f94c527e9/html5/thumbnails/3.jpg)
Precursor of miR-16 reduced contractility
Inhibitor of miR-16 increased contractility
N(rat) = 5MEAN + SEM
N(rat) = 5MEAN + SEM
p < 0.0001
p = 0.0194
17 22
16 16
Basal contractility
![Page 4: Characterization of cardiac stress (Takotsubo Cardiomyopathy)-related miRNAs ex vivo Anselm Derda, ESC Congress 2015 NHLI – Prof. Sian Harding IMTTS –](https://reader035.vdocuments.site/reader035/viewer/2022062321/56649f355503460f94c527e9/html5/thumbnails/4.jpg)
1. 2. 3.
Isolation of LV rat myocytes
Transfection & Incubation for 48h Control/Inhibitor/Mimic of miR-16
add Pertussis Toxin
Contractility/IonOptix
+ PTX
Contractility after block of Gi
![Page 5: Characterization of cardiac stress (Takotsubo Cardiomyopathy)-related miRNAs ex vivo Anselm Derda, ESC Congress 2015 NHLI – Prof. Sian Harding IMTTS –](https://reader035.vdocuments.site/reader035/viewer/2022062321/56649f355503460f94c527e9/html5/thumbnails/5.jpg)
PTX removed decreasing effect of mimic of miR-16
N(rat) = 4MEAN + SEM
p = 0.53
15 20
Contractility after block of Gi
![Page 6: Characterization of cardiac stress (Takotsubo Cardiomyopathy)-related miRNAs ex vivo Anselm Derda, ESC Congress 2015 NHLI – Prof. Sian Harding IMTTS –](https://reader035.vdocuments.site/reader035/viewer/2022062321/56649f355503460f94c527e9/html5/thumbnails/6.jpg)
1. 2. 3.
Isolation of LV rat myocytes
Transfection & Incubation for 48h Control/Inhibitor/Mimic of miR-16
Contractility/IonOptix
Contractility with isoprenaline simulating stress
+ isoprenaline
β1 β2
+ CGP
![Page 7: Characterization of cardiac stress (Takotsubo Cardiomyopathy)-related miRNAs ex vivo Anselm Derda, ESC Congress 2015 NHLI – Prof. Sian Harding IMTTS –](https://reader035.vdocuments.site/reader035/viewer/2022062321/56649f355503460f94c527e9/html5/thumbnails/7.jpg)
time
length
isoprenaline
CGPmax response
min response
Schematic: isoprenaline response
control
Inhibitor miR-16
max response min response
= = Inhibitor of miR-16 did not affect positive inotropic isoprenaline response
![Page 8: Characterization of cardiac stress (Takotsubo Cardiomyopathy)-related miRNAs ex vivo Anselm Derda, ESC Congress 2015 NHLI – Prof. Sian Harding IMTTS –](https://reader035.vdocuments.site/reader035/viewer/2022062321/56649f355503460f94c527e9/html5/thumbnails/8.jpg)
Conclusion
- miR-16 had an Gi-dependent negative inotropic effect on cardiomyocytes
- miR-16 did not affect the contractility of cardiomyocytes under stress
conditions induced by isoprenaline
- miR-16 is possibly influencing the tonic effect of Gi on contraction
- Further experiments are necessary for...
- ...evaluating signal cascade of miR-16 more precisely
- ...showing a possible interaction between miR-16 and adenylate cyclase